`
`CSL EXHIBIT 1043
`
`Page 1 0f21
`
`CSL V. Shire
`
`Page 1 of 21
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`CSL EXHIBIT 1043
`CSL v. Shire
`
`

`

`GLYCOBIOLOGY
`
`Editor-in-Cliief
`
`Ronald L. Schnaar, The Jolms Hopkins University.
`Ballinzorg' Mu USA
`
`Founding Editor
`_
`.
`.
`x
`,
`,
`,
`Gerald W. Halt, The Johns Hopkins University,
`Baltimore, MD, USA
`
`Associate Editor for Reviews
`
`Mark A. Lehrman, University ofTexas Southwestern Medical
`Centei; Dallas, TX, USA
`
`Associate Editors
`Antony Bacic, University ofMelbourne, Melbourne, Vic, Australia
`Anne lmbcrty, CERMAV-CNRS, Grenoble, France
`Reiji Kannagi, A iehi Cancer Center, Nagoya, Japan
`Ulf Lindahl, Uppsala University, Uppsaia, Sweden
`Salvatore J. Turco, University ofKentucky, Lexington, KX USA
`
`Editorial Board
`
`Markus Aebi, Zurich, Switzerland
`Linda G. Baum, Los Angeles, CA, USA
`C. Fred Brewer, Bronx, NY, USA
`lnka Brockhausen, Kingston, ON, Canada
`Karen J. Colley, Chicago, IL, USA
`Anthony P. Corfield, Bristol, UK
`Paul R. Crocker, Dundee, UK
`Richard D. Cummings, Atlanta, GA, USA
`Paul L. DeAngclis, Oklahoma City, OK, USA
`James W. Dennis, ’loronto, ON, Canada
`Kurt Drickamer, London, UK
`Alan D. Elbein, Little Rock, AR, USA
`Tamao Endo, 7bkyo, Japan
`Jeffery D. Esko, La Jolla, CA, USA
`Marilynn E. Etzler, Davis, CA, USA
`Ten Feizi, Harrow, UK
`Michael A.J. Ferguson, Dundee, UK
`Hudson H. Freeze, La Jolla, CA, USA
`Minoru Fukuda, La Jolla, CA, USA
`Koichi Fumkawa, Nagoya, Japan
`
`John T. Gallagher, Manchester; UK
`Rita Gerardy-Schahn, Hannoven Germany
`Rudolf Geyer, Giessen, Germany
`Sen-itiroh Hakomori, Seattle, WA, USA
`Robert
`Haltiwanger, Stony Brook, NY, USA
`Gunnar C. Hansson, Goteborg, Sweden
`Vincent C. Hascau, C[we[and OH, USA
`Carlos B. Hirscliberg, Boston, MA, USA
`Steven W. Homans, Leeds, UK
`Donald L. Jarvis, Laramie, WY, USA
`Kay-Hooi Khoo, Taipei, Taiwan
`Koji Kimata, Nagaknte, Japan
`Ken Kitajima, Nagoya. Japan
`Lena Kjellen, Uppsala, Sweden
`Stephan Ladisch, Washington, DC, USA
`Julie A, Leary, Davis, CA, USA
`Ludwig Lehle, Regensburg, Germany
`William J. Lennarz, Stony Brook, NY, USA
`Robert J. Linliardt, Troy, NY, USA
`Jamey D. Marth, La Jolla, CA, USA
`alcolm J. McConville, Parkville, Australia
`{elley W. Moremcn, Athens, GA, USA
`Barbara Mulloy, Potters Bar, UK
`.{afael Oriol, Villejnifl France
`Vlonica M. l’alcic, Copenhagen, Denmark
`Armando J. Parodi, Baenos Aires, Argentina
`James C. Paulson, La Jolla, CA. USA
`Vlichael Pierce, Athens, GA, USA
`Robert Sacksteiu, Boston, MA, USA
`~lcnrik V. Scheller, Berkeley. CA, USA
`Joel H. Shaper, Baltimore, MD, USA
`Vathan Sharon, Re/zovot, Israel
`Pamela Stanley, New York, NY, USA
`Akemi Suzuki, Hiratsaka, Japan
`\laoyuki 'l'aniguchi, Osaka, Japan
`Maureen E. Taylor, London, UK
`Gerrit van Mcer, Utrecht, The Netherlands
`AjitVarki, La Jolla, CA. USA
`Charles J. Waeehter, Lexington, KY, USA
`Kalsuko Yamashita, Tokyo, Japan
`Masaki Yanagishita, Tokyo, Japan
`
`
`
`Front cover: Muscles from ma’x mice, a model for human Duchenne Muscular Dystrophy, exhibit increased expression ol‘galectin—l dur-
`ing degeneration. Sections oi‘diaphragm muscle from control (left panels) and mdx mice (right panels) at 4 weeks of age (top panels) and
`6 weeks of age (bottom panels) are shown. Sections were stained with anti-galectin—l (green) and DAPI (nuclei, blue). Non-peripherally
`nucleated myofibers, detected only in "if/X muscles, are indicated by arrowheads. Bar = 10 um. For further details, see: Ccrri DG, Rodrigues
`LC, Stowcll SR, Araujo DD, Coelho MC, Oliveira SR, Bizario JC, Cummings RD, Dias—Baruffi M, and Costa MC. 2008. Degeneration ol
`dystrophic or injured skeletal muscles induces high expression of Galcctin-l. lecobiology. 18:842—850.
`
`Page 2 of21
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`Page 2 of 21
`
`

`

`CLOSING REMARKS & RECEPTION
`
`
` 8:30 —10:00 am
`
`ANNUAL CONFERENCE
`of the
`SOCIETY FOR GLYCOBIOLOGY
`November 12 — 15, 2009
`Catamaran Resort Hotel, San Diego, CA
`
`eociety fol.
`
`I,”
`
`‘
`
`' (5,,“
`
`.\
`yCo b io\°°’
`
`c l
`
`CONFERENCE LOCATION. All technical sessions, posters and exhibits will be at the Catamaran Resort Hotel, 3999
`Mission Blvd, San Diego, CA 92109. The conference rate is $159 for single/double. Please refer to www.glycobiology.org
`for more detailed inlbrmation.
`DEADLINES
`
`Friday, July 24
`
`Friday, August 14
`Friday, August 14
`Friday, October 2
`Friday, October 9
`
`Submission of nominations for the Karl Meyer Award
`and Rosalind Kornfeld Award
`Submission of abstracts
`
`Submission of applications for Student Travel Awards
`Advance conference registration
`Reservations at the Catamaran Resort Hotel
`
`ORGANIZER
`
`Thomas N Oeltm a n n , I’residenl and Meeting Chair
`Vanderbilt University, School of Medicine, Nashville, TN= presidentfiilglycobiologvere or
`Lonmeltinann@vanderbiltcdu
`PRELIMINARY PROGRAM
`
`8:00 am — 5:30 pm
`
`SATELLITE MEETING: GLYCANS IN CELL COMMUNICATION
`
`Annual meeting ofthe Consortium for Functional Glycomics Participating Investigators
`Attendance is open to all interested scientists and isfree ofc'lmi‘ge.
`Go to http://glvcomics.scrigpsedu/Pl2009.htinl for program information
`Please notify Anna Crie (annacrie@scripps.edu) before October 9
`SATELLITE SYMPOSIUM: ADVANCES IN GLYCOPROTEIN TECHNOLOGIES
`
`8:30 am 4 1:00 pm
`
`Organized by Qun Zhou, Genzyme Corp, Cambridge, MA and Samnang Tep, Biogen ldec,
`Cambridge, MA,
`$40
`
`CONFERENCE OPENING
`
`Opening Remarks: Thomas N Oeltmann, President, Societyfm' Glycobio/ogy
`SESSION I: GLYCOBIOLOGY OF THE IMMUNE SYSTEM
`KORNFELD AWARD AND LECTURE
`WELCOME RECEPTION
`
`NnV—t
`
`aw
`U)
`'52
`;‘E50
`=>
`H94
`
`7:00 a 7:10 pm
`
`7: 10 ~ 8:30 pm
`
`8:30 — 9:00 pm
`9:00— 10:00pm
`
`v,Fridav‘November13
`
`Saturday,
`
`November14
`
`Sunday,November15
`
`8:30 - 10:00am
`
`10:30am 12:20 pm
`
`2:00 4 4:00pm
`
`SESSION II: GLYCOBIOLOGY OF DISEASE
`SESSION III: GLYCOBIOLOGY IN DEVELOPMENT
`POSTERS and EXHIBITS
`
`SESSION IV: GLYCOBIOLOGY OF RECOGNITION: PROTEIN-
`4:00 7 5:50 pm
`CARBOHYDRATEINTERACTIONS
`
`SESSION V: PATHOG EN GLYCOBIOLOGY
`
`10:30am — 12:20 pm
`
`2:00 — 4:00 pm
`4:00 4 4:30 pm
`
`4:45 — 5:30 pm
`7:00 — 9:30pm
`
`SESSION VI: ASMB Guest Society Session: GLYCOBIOLOGY AND THE
`EXTRACELLULAR MATRIX
`
`POSTERS and EXHIBITS
`
`BUSINESS MEETING
`
`KARL MEYER AWARD AND LECTURE
`
`BANQUET. Nominal fee. Extra tickets for guests may be ordered.
`
`8:304 10:00 am
`
`SESSION VII: GLYCOBIOLOGY AND THE NERVOUS SYSTEM
`
`10:30 am — 12:20 pm
`
`2:00 — 4:00 pm
`
`4:00 ~ 5:40 pm
`
`5:40 — 6:00 pm
`
`SESSION VIII: CYTOPLASMIC GLYCOSYLATION, METABOLISM, AND O—
`GLCNAC MODIFICATION
`
`POSTERS and EXHIBITS
`
`SESSION IX: GLYCAN RECOGNITION IN THE ER
`
`
`
`Page 3 of 21
`
`

`

`DEADLINE: Friday, October 2
`ADVANCE REGISTRATION FORM
`ADVANCE REGISTER BY MAIL, FAX, OR ONLINE. You may advance register by mailing or faxing this page (see
`below) OR on-line at wwwglycobiology.org. The deadline for advance registration is October 7 7009.
`
`ON SITE REGISTRATION. After October 2, please register on site. On site registration fees will be $50 more than advance
`registration. On site registration begins at 3 pm, Thursday, November [2 at the Catamaran Resort Hotel.
`
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`SATELLITE SYMPOSIUM (8:30 am 7 1:00 pm, Thursday, November 12, $40)
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`Enter $10 for the banquet $
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`Page 4 of21
`
`Page 4 of 21
`
`

`

`Mutational and functional analysis of Large in a novel
`CHO glycosylation mutant
`Establishment of a real-time analytical method for free
`oligosaecharide transport from the ER to the cytosol
`Catabolism of flocculosin, an antimicrobial metabolite
`produced by I’seudozymaflocculosa
`Production of human B-hexosaminidase A with highly
`phosphorylated N-glycans by the overexpression of the
`()gamca minuta MNN4 gene
`'IWvo distinct oc-L-fucosidases from Bifidobacterium
`bifidum are essential for the utilization of fucosylated
`milk oligosaccharides and glycoconjugates
`
`1046
`
`Glycolnology
`Volume l9 Number 9 September 200‘)
`
`Contents
`
`GLYCO—FORUM SECTION
`
`Meeting Announcements
`
`REVIEW
`
`’
`
`.
`
`,
`
`OXFORD *1
`
`‘
`
`y
`
`'
`
`Optimal and consistent protein glycosylation in
`mammalian cell culture
`
`P I-Iossler, SF Khattak, and Z] Li
`
`COMMUNICATION
`
`The specific localization of seminolipid molecular
`species on mouse testis during testicular maturation
`revealed by imaging mass spectrometry
`
`ORIGINAL ARTICLES
`
`N Goto—Inoue, T Hayasaka, N Zaima, and M Seton
`
`Glycosylation profiles of epitope-specific anti-B-amyloid
`antibodies revealed by liquid chromatography—mass
`spectrometry
`
`I Perdivara, LJ Delerding, C Cozma, KB Tomer, and
`M Przybylski
`
`JT Aguilan, S Sundaram, E Nieves, and P Stanley
`
`Y Haga, K Totani, Y Ito, and T Suzuki
`
`B Mimee, C Labbé, and RR Be’langer
`
`H Akeboshi, Y Kasaharn, D Tsuji, K Itoh, H Sakuraba,
`Y Chiba, and Y Jigami
`
`H Ashida, A Miyake, M Kiyohara, J Wada, E Yoshida,
`H Kumagai, T Katayama, and K Yamamoto
`
`Unusual accumulation of sull'ated glycosphingolipids in
`colon cancer cells
`
`K Shida, Y Misonou, H Korekane, Y Seki, S Noura,
`M Ohue, K Honke, and Y Miyamoto
`
`The ot-galactomannan Davanat binds galectin-l at a site
`different from the conventional galectin carbohydrate
`binding domain
`
`Heterodisaccharide 4-0-(N-acetyl-B-D-glucosaminyl)-
`I)-glueosamine is a specific inducer of chitinolytic
`enzyme production in Vibrios harboring chitin
`oligosaceharide deacetylase genes
`
`MC Miller, A Klyosov, and KH Mayo
`
`T Hirano, K Kadokura, T Ikegami, Y Shigeta, Y Kumaki,
`W Hakamata, T Oku, and T Nishio
`
`Page 5 of 21
`
`

`

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`Page 6 of 21
`
`

`

`Glycobiology vol. 19m), 9 pl 935, 2009
`doi: 10‘ [DOB/glycob/cwp 108
`
`Glyco—Forum section
`
`
`
`Meeting Announcements
`
`6*" International Conference on
`
`Proteoglycans
`
`September 13—17, 2009, Aix-les-Bains, France
`
`Conference eo-organizers:
`H. Lortat-Jacob (Grenoble, France)
`J. Van den Born (Gronii’igen, The Netherlands)
`Contact: contact.pg2009@ibs.fr
`
`For details please visit: http://pg2009-france.ibs.fr/.
`
`2009 Annual Meeting of the Society for
`Glycobiology
`
`November 12—15, 2009, Catamaran Hotel, San Diego,
`California
`
`Program Chair: Tom Oeltmaun, Vanderbilt University
`For details please visit http://www.glycobiology.org
`
`20‘“ International Symposium on
`Glycoconjugates
`Caribe Hilton Hotel, San Juan, Puerto Rico
`November 29—December 4, 2009
`
`For further details, please visit http://www.glyco20.org.
`
`Dr. Dipak K. Banerjee
`Department of Biochemistry
`School of Medicine, University of Puerto Rico
`Medical Sciences Campus
`GPO Box 365067, San Juan, PR 00936—5067
`Telephone: +1-787—758-2525 ext. 1624 or +1-787-758-7090
`Fax: +1-787-274-8724
`
`E—mail: dbanerjee@rcm.upredu (Scientific Secretariat) or
`ceast‘aneda@leemanagemcntcom (Organizational Secretariat)
`
`Published by Oxford University Press 200‘).
`
`Page 7 of21
`
`935
`
`Page 7 of 21
`
`

`

`Glycobiology vol. 19 no. 9 pp. 936-949, 2009
`doi:10.1093/g1ycob/cwp079
`Advance Access publication on June 3, 2009
`
`REVIEW
`
`Optimal and consistent protein glycosylation in mammalian cell culture
`
`Patrick HosslerM’Z, Sar 'at F Khattaks", and
`Zheng Jian Li”
`
`4Process Sciences Upstream, Technical Operations, Bristol-Myers Squibb
`Company, East Syracuse, NY 13057, USA
`
`Received UNA/nil 6, 2009; wvisecl on June I, 2009; accepted on Jth I, 200‘)
`
`In the biopharmaceutical industry, mammalian cell culture
`systems, especially Chinese hamster ovary (CHO) cells, are
`predominantly used for the production oftherapeutic glyco-
`proteins. Glycosylation is a critical protein quality attribute
`that can modulate the efficacy of a commercial therapeu-
`tic glycoprotein. Obtaining a consistent glycoform profile in
`production is desired due to regulatory concerns because a
`molecule can be defined by its carbohydrate structures. An
`optimal profile may involve a spectrum of product glycans
`that confers a desired therapeutic efficacy, or a homogeneous
`glycoform profile that can be systemically screened for. Stud-
`ies have shown some degree of protein glycosylation control
`in mammalian cell culture, through cellular, media, and pro-
`cess effects. Studies upon our own bioprocesses to produce
`fusion proteins and monoclonal antibodies have shown an
`intricate relationship between these variables and the result-
`ing protein quality. Glycosylation optimization will improve
`therapeutic efficacy and is an ongoing goal for researchers
`in academia and industry alike. This review will focus on
`the advancements made in glycosylation control in a manu-
`facturing process, as well as the next steps in understanding
`and controlling protein glycosylation.
`
`Keywords: bioprocessing/mammalian cell culture/protein
`glycosylation/protein therapeutics/protein quality
`
`Introduction
`
`Protein glycosylation is of paramount importance to the efficacy
`and manufacturing of therapeutic glycoproteins. Mammalian
`cell expression systems are the preferred method for the com-
`mercial production of these glycoproteins because their innate
`protein processing machinery, including that of protein glyco—
`sylation, closely resembles that in human.
`Glycosylation of proteins takes on the form of oligosaccha—
`rides attached to either the side chain of asparagine (N—linked)
`or serine/threonine (0-linked) with the former being the most
`
`
`ITo whom correspondence should be addressed: Tel: el— 1-315-432-2131 ; e-mail:
`zliengjian.li@bms.com
`2Present address: Abbott Biorcsearch Center, Abbot Labor
`MA 01605, USA,
`3These authors contributed equally to this work.
`
`alorles, Worcester,
`
`prominent (Warren 1993; Helenius and Aebi 2001; Sinclair and
`Elliott 2005). Glycans have a very prominent role toward af-
`fecting therapeutic efficacy and determining the in vivo half-life
`(Elliott ct al. 2003). It is due to both of these features that the
`glycoform profile of a therapeutic glycoprotein must be exten-
`sivel y characterized in order to meet regulatory agency demands
`(FDA 1996).
`In this review, we summarize the salient features of protein
`glycosylation control from a bioprocess perspective. Through
`our own process development and characterizatimt efforts with
`fusion proteins such as OrenciaTM and Belatacept and motio-
`clonal antibody Ipilimumab, we have gained an appreciation
`over the past, present, and future state of the control of this
`very important metabolic pathway. Table I lists glycosylated
`therapeutics currently on the market, which is the impetus
`for understanding their metabolic control. Although CHO cells
`are the most prevalent for producing glycoprotein therapeutics,
`other cell lines such as human embryonic kidney cells (HEK)
`(Durocher ct al. 2007), baby hamster kidney (BHK), mouse
`myeloma (N50), and human retinal cells (PERC.6)
`(Jones
`et al. 2003; Wurm 2004; Petricciani and Sheets 2008) are being
`developed as high producing cell lines.
`There have been numerous reports on the effects of cell type,
`culture process, and culture media toward the resulting glyco—
`form profile. This review will summarize these efforts, as well
`as new attempts toward achieving uniform, consistent, and/or
`optimal glycosylation. These efforts share a common need for
`a more fundamental understanding of the metabolic pathway,
`which is the next logical progression from the current paradigm
`of glycosylation maintenance. The ultimate goal would be to
`take the lessons learned from previous studies, combined with
`new technologies, to transform a well-characterized cell line to-
`ward one that can control the glycosylation profile. In order to
`truly incorporate process quality by design (QbD) into the final
`glycoprotein product (Rathore and Winkle 2009), more funda—
`mental studies on the dependence of glycosylation on process
`conditions and cellular physiology will be required to gain phys-
`iological insight. The information gathered from such research
`together with clinical studies will continue to enhance our capa-
`bility toward generating higth efficacious glycoprotein drugs.
`
`Protein glycosylation and product quality
`
`Metabolic pathway
`The addition of oligosaccharides onto a glycoprotein is a com-
`plex metabolic pathway, characterized by the en bloc transfer
`of polysaccharide chains, as well as the step-wise addition and
`removal of individual monosaccharides. The number of palli-
`ways traversed is dependent on reaction site accessibility, the
`
`© The Author 2009. Published by Oxford University Press All rights reserved. For permissions, please c-mail: journalspermissi0ns@oxterdjournals.org
`
`936
`
`Page 8 of21
`
`Page 8 of 21
`
`

`

`Optimal protein glycosylation in mammalian cell culture
`
`
`
`Product
`
`Class
`
`Mode of action
`
`Indication
`
`Cell line
`
`Company
`
`Table I. Glycosylated therapeutics approved by the FDA and/or EMEA
`
`Atanesp
`(Darbepoetin
`alfa)
`Arcalyst
`(Rilottaccpt)
`Avastin
`(Bevacizumab)
`Avonex
`(Interferon
`['5— 1 it)
`Cerczymc
`(Imigluccrasc)
`
`Erythropoicsis
`stimulating
`protein
`IL-l Trap
`
`rMab
`
`Interferon
`
`Enzyme
`
`Elaprase
`(lduronate
`sulfatase)
`
`Enbrel
`(Etanerccpt)
`
`Epogcn (tipoetin
`alfa)
`
`Enzyme
`
`Fusion protein
`
`Erythropoiesis
`stimulating
`protein
`
`Erbiqu
`(Cctuximab)
`
`rMab
`
`Regulates red blood cell production
`
`Anemia
`
`Binds lL-li‘i to prevent the interaction to cell surface
`receptors
`Binds to the vascular cndothelial growth factor
`(VEGF) to inhibit angiogencsis
`Binds to type I interferon] receptors to activate two
`.lak tyrosine kinases
`
`(.‘ryoporin-associatcd
`periodic syndromes
`Colorectal cancer
`
`CHO
`
`CHO
`
`CHO
`
`Amgcn
`
`Regeneron
`
`Genentech
`
`Multiple sclerosis (MS)
`
`Cl l()
`
`Biogcn Idcc
`
`Gaucher Disease
`
`Engineered to ltavc mannose-tcrtninatcd
`oligosaccharidc chains that are recognized by
`endocytic carbohydrate receptors on
`macrophages. Catalyzes hydrolysis of glycolipid
`glucocerebrosidc to glucose and ceramide in
`those macrophages which accumulate lipids in
`Gaucher disease
`lIydt‘olyzcs the 2-sulfatc esters of terminal iduronate Hunter syndrome
`sulfate residues from the glycosaminoglycans
`dcrmatan stllfate and heparan sulfate in the
`lysosomes of various cell types
`Mimics inhibitory effects of naturally occurring
`soluble TNF receptors to reduce inflammatory
`I'CSPOlth
`Recombinant human erythropoietin interacts with
`cryt tt'opoietin (EPO) receptors to stimulate
`production of red blood cells from bone marrow
`stem cells
`
`Rheumatoid arthritis
`
`Anemia
`
`CIIO
`
`Genzyme
`
`HT-IOSO
`
`Shire
`
`CHO
`
`CllO
`
`Amgcn
`
`Amgcn, Kirin
`
`Colorcclal cancer
`
`SI’Z/O
`
`lmdon‘3/BMS
`
`GCHCHYCCh
`
`IIcrccptin
`('l'rastuzumab)
`Wellferon
`(Interferon u)
`
`rMab
`
`Cytokine
`
`Mirccra (Mcthoxy
`polyethylene
`glycol-cpoctin
`beta)
`Myozymc
`(Alglucosidase
`alfa)
`Naglazyme
`(Galsttlfase)
`
`NeoReeormon
`(Epoetin beta)
`
`Orencia
`(Abataccpt)
`
`l’rocrit/Eprcx
`(Epoctin alfa)
`
`Erytht‘opoiesis
`stimulating
`protein
`
`Enzyme
`
`Enzyme
`
`lirythropoicsis
`stimulating
`protein
`
`Fusion Protein
`
`Erythropoiesis
`stimulating
`protein
`
`Rebif (Interferon
`p-13)
`
`Cytokinc
`
`rMab
`
`
`
`Binds o the extracellular domain of the epidermal
`growth factor (EGFR) preventing activation of
`EGFR to impair cell growth and proliferation
`Binds o IIER2+ tumor cells, blocks downstream
`HERIZ signaling to inhibit proliferation of cells
`Binds o type I interferon receptors (IFNARI and
`lFNARZc) which, upon dimerization, activate two
`Jak Janus kinase) tyrosine kinases (.Iakl and
`'l'yk2). Uprcgulates expression of MHC 1 proteins
`Recotn inant human erythropoictin interacts with
`etyt iropoietin (EPO) receptors to stimulate
`prot action of red blood cells from bone marrow
`stem cells
`RecomJinant acid a«glucosidasc (GAA) to replace
`GAA deficiencies
`
`
`
`Recont iinant form of polymorphic human enzyme
`Niacetylgalactosamino 4—sullatasc which
`catabolizes glycosaininoglycans (GAG)
`Recom inaltt human crythropoietin interacts with
`crythropoictin (EI’O) receptors to stimulate
`production of red blood cells from bone marrow
`stem cells
`C'I‘LAL -lg acts as a selective modulator of the
`costimulatory signal required for full 'I'—cell
`activation
`Recombinant human erythropoictin interacts with
`crythropoietin (EI’O) receptors to stimulate
`production of red blood cells from bone marrow
`stein cells
`Immunotnodulation through the induction of cell
`membrane components of the major
`histocompatibility complex
`Binds to ’I‘NF and inhibits ’I‘NF action
`
`Breast cancer
`
`CHO
`
`Chronic hepatitis C
`
`I'luman
`lympho-
`blastoid
`
`Wellcome
`
`Anemia
`
`Pompe disease
`
`Mucopolysaccharidosis
`VI (MI‘S Vl)
`
`Anemia
`
`CHO
`
`CHO
`
`CHO
`
`CHO
`
`Rheumatoid arthritis
`
`CHO
`
`Anemia
`
`CHO
`
`Multiple sclerosis (MS)
`
`CHO
`
`Hoffmann-La
`Roche
`
`Genzyme
`
`Biomarin
`
`Roche
`
`Bristol-Myers
`Squibb
`
`10"“5011 &
`JOIIHSOIL
`SChering'
`I’lmlgh
`Merck Scrono
`
`SI’Z/O
`
`Crohn’s disease &
`Rcmieade
`Centocor/Johnson
`Rheumatoid arthritis
`(Inmxilllilb)
`& Johnson
`Non-I'lodgkins
`Binds to the cluster of dilfcrcntiation 20 (CD20)
`Rituxan
`rMab
`CHO
`Gcncntcch (IDEC)
`lymphoma
`which is expressed on B-cells. Fc portion
`(Rituximah)
`mediates ADCC and CDC
`
`Page 9 of21
`
`(Continued)
`
`937
`
`Page 9 of 21
`
`

`

`l’ Hossler et al.
`
`Table 1. (Continued)
`
`Product
`Class
`Mode of action
`
`
` Indication Cell line Company
`
`
`
`Soliris
`(Eculizumab)
`
`rMab
`
`Synagis
`(Palivizumab)
`TNKase
`(Tenecteplase)
`(IPA)
`
`Tysabri
`(Natalizumab)
`
`Vectibix
`(Panitumumab)
`
`chapax
`(Daclizumab)
`
`rMab
`
`Enzyme
`
`rMab
`
`rMab
`
`rMab
`
`Binds to the complement protein C5, inhibiting
`terminal complement mediated intravascular
`hcmolysis
`Targets an epitope in the A antigenic site of the F
`protein of respiratory syncytial virus (RSV)
`Recombinant fibrin—specific plasminogen activator
`
`Paroxysmal nocturnal
`ltemoglobinuria
`
`Respiratory syncytial
`virus
`Acute myocardial
`infarction
`
`C110
`
`NSO
`
`CHO
`
`Alcxion
`
`Medlminune
`
`(icnctech
`
`Binds to 014 integrin of adhesion molecule VLA-4
`and sterically inhibits binding of VLA-4 to
`VCAM~1
`
`Multiple sclerosis (MS)
`
`CHO
`
`Biogen ldec
`
`Binds to EGFR
`
`Binds to the OL subunit (p.55 01, CD25, or Tac subunit)
`of human lL—2 receptor expressed on the surface
`of activated lymphocytes
`
`Colorcctal carcinoma
`
`Acute organ rejection
`
`CHO
`
`Sl’2/(l
`
`Amgen
`
`l'loffmann-La
`Roche
`
`enzymatic substrate specificities, as well as spatial localiza—
`tion of the various enzymes and nucleotide—sugar substrates that
`are necessary for the reactions to proceed in a particular order
`(Krambeck and Betenbaugh 2005; Butler 2006).
`Whereas N—linked glycosylation initiates primarily in the ER
`of mammalian cells, 0~linked glycosylation has been shown
`to initiate in either the ER or Golgi apparatus. Typical N— and
`O-glycan structures with their symbolic monosaccharide repre-
`sentation and nomenclature are shown in Figure 1.
`There exists the potential for a diverse array of product gly-
`cans on each glycoprotein molecule as demonstrated in the
`KEGG GLYCAN database (Hashimoto et al. 2006). In most
`industrial bioprocesses, this large repertoire of glycans is typi—
`cally not seen on recombinant glycoproteins expressed by mam-
`malian cells such as CH0 and N80, suggesting that there may
`be pathway constraints. Control of this metabolic pathway can
`be manifested through a variety of different conditions, includ—
`ing the cell lines, bioprocess, and cell culture media used for
`protein production.
`
`Mormsaccharide and oligosaccltaride roles in protein
`therapeutic ejficacy
`
`N— and O-linked glycans have been shown to have a large effect
`on the immunogenicity, efficacy. solubility, and half-life ofcom-
`mercial biologics (Lis and Sharon 1993; Van den Steen et al.
`1998; Lowe and Marth 2003). Individual monosaccharidcs on
`N— and O-glycan structures can help determine a glycoprotein’s
`therapeutic efficacy by their immunological and pharmacoki-
`netic impact (Sethuraman and Stadheim 2006).
`
`N—Ijn/(erl Glycosylation. Figure 2 is a schematic of N—linked
`glycosylation for a protein therapeutic in a CHO cell culture.
`The terminal N—acetylneuraminic acid (NANA) content in a
`glycoprotein has been shown in many cases to be intricately
`linked to circulatory half-life, and thus is a typical marker for
`product quality assessment in protein therapeutics due to its
`proven role toward affecting circulatory half-life. Exceptions
`have been shown with monoclonal IgGl antibodies, where there
`was no significant difference in the clearance rate of molecules
`carrying different amounts of sialic acid (